Re: Mixing primary and secondary before staining?

From:

"Connolly, Brett M"

This was described in a paper by Gu et al. in 1995. Cell Vision 2(1), 1995
pp, 52-58
Brett
Brett M. Connolly, Ph.D.
Merck & Co.,Inc.
MRL, Imaging
WP26A-3000
PO Box 4
West Point, PA 19486
PH 215-652-2501
fax. 215-652-2075
e-mail. brett_connolly@merck.com
-----Original Message-----
From: Ed Boyden [mailto:ed_boyden@hotmail.com]
Sent: Tuesday, April 29, 2003 2:35 PM
To: histonet@pathology.swmed.edu
Subject: Mixing primary and secondary before staining?
Dear All,
I have a large number of brain samples to process for immunocytochemistry
with a primary and then a fluorescent secondary antibody. I was wondering if
it would be possible to save steps by premixing the primary and fluorescent
secondary together, then staining with the mix and doing washes like normal.
Blocking with serum would preceded the staining, and thus should prevent any
nonspecific background. Also, since only the primary and secondary are
present when they are binding to each other, this might actually reduce
nonspecific binding to parts of the tissue.
Any ideas? I have not heard of anyone doing this before, but there were no
contraindications either listed in the histonet archives.
Best,
Ed
ed_boyden@hotmail.com
Dept. of Molecular and Cellular Physiology
Stanford Neuroscience Program
Stanford, CA 94305-5345
phone (650) 725-7564
fax (650) 725-8021
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